Cystic Fibrosis (CF) is a lethal cureless disease affecting more than 30,000 children and adults in the US, caused by a single-gene defect that leads to impaired function of the CFTR protein responsible for chloride ion transport. The pathological hallmark of CF is the formation of thick mucus lining the lung airways, which cannot be promptly cleared by ciliary action. The accumulated mucus obstructs the lungs, makes breathing difficult and promotes bacterial invasion, ultimately eliciting a massive inflammatory response. Furthermore, the mucosal layer, which is the primary extracellular barrier in the CF lung, is a major cause for the failure of varied therapeutic approaches to CF. In particular, the thick mucus has been a critical obstacle for aerosol gene therapy for CF (i.e. delivery of gene vectors through the lung airways rather than intravenously), which generally holds tremendous potential as a therapeutic avenue. The goal of the proposed research is the construction and testing of a new nanoparticle-based platform for CF gene therapy that would facilitate penetration through the lung mucosal barrier, leading to overall higher transfection efficiency. The laboratory of Prof. Justin Hanes, Head of Therapeutics at the Institute of NanoBioTechnology at Johns Hopkins University (the host researcher), has been in the scientific and technological forefront for developing innovative polymeric nanoparticle (NP) carriers for pulmonary therapeutics. Specifically, the primary vehicle pursued in the research will be porous polymeric NPs modulated with polyethylene-glycol (PEG), recently shown in the Hanes laboratory to significantly enhance mucosal permeation. Overall, the proposed work is guided by the hypothesis that a comprehensive and quantitative approach to the development of polymeric NP assemblies exhibiting enhanced penetration through mucosal barriers is essential, and indeed possible, for successful CF gene therapy. The conceptual and technological advances pursued in the proposed project could open the way for varied therapeutic avenues for pulmonary diseases and targeted drug delivery through physiological aerial pathways. PUBLIC HEALTH RELEVANCE: The proposed research aims to develop an innovative gene therapy approach for cystic fibrosis, a cureless lethal lung disease often leading to considerable human suffering. The research will focus on a new technology for effective delivery of "gene nano-carriers" through the thick mucus barrier in CF lungs.